|Publication number||US5733542 A|
|Application number||US 08/377,771|
|Publication date||31 Mar 1998|
|Filing date||24 Jan 1995|
|Priority date||16 Nov 1990|
|Also published as||WO1996023058A1|
|Publication number||08377771, 377771, US 5733542 A, US 5733542A, US-A-5733542, US5733542 A, US5733542A|
|Inventors||Stephen E. Haynesworth, Arnold I. Caplan, Stanton L. Gerson, Hillard M. Lazarus|
|Original Assignee||Haynesworth; Stephen E., Caplan; Arnold I., Gerson; Stanton L., Lazarus; Hillard M.|
|Export Citation||BiBTeX, EndNote, RefMan|
|Patent Citations (2), Non-Patent Citations (2), Referenced by (171), Classifications (39), Legal Events (7)|
|External Links: USPTO, USPTO Assignment, Espacenet|
This is a continuation-in-part of U.S. Ser. No. 08/193,262, filed Feb. 8, 1994 (U.S. Pat. No. 5,486,359 issued on Jan. 23, 1996), which is a continuation-in-part of U.S. Ser. No. 08/034,272, filed Mar. 22, 1993 (abandoned), which is a continuation-in-part of U.S. Ser. No. 07/716,917 filed Jun. 18, 1991 (abandoned), which is a continuation-in-part of U.S. Ser. No. 07/615,430 filed Nov. 16, 1990 (abandoned); and a continuation-in-part of U.S. Ser. No. 08/038,517, filed Mar. 29, 1993 (abandoned), which is a divisional of U.S. Ser. No. 07/614,912, filed Nov. 16, 1990 (U.S. Pat. No. 5,226,914 issued on Jul. 13, 1993); and a continuation-in-part of U.S. Ser. No. 08/038,512, filed Mar. 29, 1993 (abandoned), which is a divisional of 07/614,915, filed Nov. 16, 1990 (U.S. Pat. No. 5,197,985 issued on Mar. 30, 1993).
The present invention is directed to various methods and devices for using mesenchymal stem cells (MSCs) to enhance bone marrow engraftment. Mesenchymal stem cells are the formative pluripotent blast cells found in the bone that are capable of differentiating into any of the specific types of connective tissues (i.e., the tissues of the adipose, areolar, osseous, cartilaginous, elastic, and fibrous connective tissues) depending upon various environmental influences. Although these cells are normally present at very low frequencies in bone marrow and other mesenchymal tissues, the inventors of the present invention have discovered a process for isolating, purifying, and greatly replicating the mesenchymal stem cells in culture, i.e. in vitro. This discovery is the subject of co-pending U.S. patent application Ser. No. 193,262, filed Feb. 8, 1994. The methods and devices of the invention utilize such isolated and culture-expanded mesenchymal stem cells (MSCs). Under selected conditions, they can be induced to differentiate into different types of skeletal and connective tissues such as bone, cartilage, tendon, ligament, muscle, other connective tissues and marrow stroma.
The marrow stroma provides the scaffolding as well as soluble factors which direct and support blood cell synthesis, i.e., hematopoiesis. The present invention is directed to a method to improve the process of bone marrow transplantation which is used to regenerate blood cells and marrow tissue in patients where their marrow is depleted or destroyed, such as, for example, during intensive radiation and chemotherapy treatment. Along this line, the inventors have discovered that, after lethal doses of radiation, culture-expanded MSCs can increase survival and decrease the time of blood and marrow cell regeneration when transplanted with complete bone marrow.
The present invention provides a method for enhancing the regeneration of marrow tissue through improved bone marrow transplantation using MSCs. The method for enhancing bone marrow engraftment comprises administering to an individual in need thereof, (i) mesenchymal stem cells and (ii) a bone marrow graft, wherein said mesenchymal stem cells are administered in an amount effective to promote engraftment of the bone marrow in the individual. More particularly, one embodiment of the invention is directed to a method for using a culture medium comprised of Dulbecco's Modified Essential Medium with low glucose (DMEM-LG) or medium 199 plus 1% human albumin as a vehicle or carrier for MSCs which, when administered systemically, will migrate, or home, to the marrow cavity and differentiate into marrow stroma, thereby regenerating the marrow stroma. The MSCs can be administered systemically, e.g., intravenously, into various delivery sites or directly into the bone.
A further aspect of the present invention is directed to the timing of injection of the MSCs into the patient relative to the bone marrow transplantation. In one embodiment, the MSCs are injected simultaneously with the bone marrow. In another embodiment, the MSCs are administered before or after the injection of bone marrow.
The present invention is useful to enhance the effectiveness of bone marrow transplantation as a treatment for cancer. The treatment of cancer by x-irradiation or alkylating therapy destroys the bone marrow microenvironment as well as the hematopoietic stem cells. The current treatment is to transplant the patient after marrow ablation with bone marrow which has been previously harvested and cryopreserved. However, because the bone marrow microenvironment is destroyed, bone marrow engraftment is delayed until the stromal environment is restored. As a result, a critical aspect of the present invention is directed to the advantages of transplanting isolated, purified, culture-expanded MSCs to accelerate the process of stromal reconstitution and ultimately marrow engraftment.
Modes of administration of the MSC preparation include but are not limited to systemic intravenous injection and injection directly to the intended site of activity. The preparation can be administered by any convenient route, for example by infusion or bolus injection and can be administered together with other biologically active agents. Administration is preferably systemic.
The present invention also provides pharmaceutical compositions. Such compositions comprise a therapeutically effective amount of the MSCs, and a pharmaceutically acceptable carrier or excipient. Such a carrier includes but is not limited to medium 199 plus 1% serum albumin, saline, buffered saline, dextrose, water, and combinations thereof. The formulation should suit the mode of administration.
In a preferred embodiment, the MSC preparation or composition is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous administration to human beings. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. Where necessary, the composition may also include a local anesthetic to ameliorate any pain at the site of the injection. Generally, the ingredients are supplied either separately or mixed together in unit dosage form, for example, as a cryopreserved concentrate in a hermetically sealed container such as an ampoule indicating the quantity of active agent. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to administration.
The method of the invention can be altered, particularly by (1) increasing or decreasing the time interval between injecting MSCs and implanting the tissue; (2) increasing or decreasing the amount of MSCs injected; (3) varying the number of MSC injections; (4) varying the method of delivery of MSC; or (5) varying the source of MSCs. Although MSCs derived from the tissue donor is preferable, the MSCs can be obtained from other individuals or species, or from genetically-engineered inbred donor strains, or from in vitro cell culture.
The MSC preparation is used in an amount effective to promote engraftment of bone marrow in the recipient. In general, such amount is at least 1×104 MSC per kg of body weight and most generally need not be more than 7×105 MSC/kg. Preferably, it is at least about 2×105 MSC/kg prior to graft introduction and usually need not be more than about 7×105 MSC/kg. The MSC preparation may be administered concurrently with the bone marrow transplant or for a period prior to graft introduction of at least about 7 days but generally not to exceed 30 days, with a typical therapeutic treatment period of 7 to 14 days. The MSC preparation preferably is administered either intravenously one to three times per day, and may be adjusted to meet optimal efficacy and pharmacological dosing.
The invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical preparation of the invention. Associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration.
Marrow in femoral head cancellous bone pieces was obtained from patients with degenerative joint disease during hip or knee joint replacement surgery. In addition, marrow was also obtained by iliac aspirate from normal donors and oncology patients who were having marrow harvested for future bone marrow transplantation. All of the oncology patients had malignancies unrelated to the stromal cells and the stromal cells expressed normal karyotype.
Marrow for cell culture was prepared from plugs of cancellous bone marrow as follows. Plugs of cancellous bone marrow (0.5-1.5 ml) were transferred to sterile tubes to which 25 ml Dulbecco's Modified Eagles Medium with low glucose (DMEM-LG) (GIBCO, Grand Island, N.Y.) with selected batches of 10% fetal bovine serum (complete medium) was added. The tubes were vortexed to disperse the marrow, then spun at 1000 rpm for 5 minutes to pellet cells and bone pieces. The supernatant and fat layer were removed and the marrow and bone were reconstituted in 5 ml complete medium and vortexed to suspend the marrow cells. The suspended cells were collected with a 10 ml syringe fitted with a 16 gauge needle and transferred to separate tubes. Bone pieces were reconstituted in 5 ml of complete medium and the marrow cells were collected as before. Collection of marrow cells was considered complete when a pellet of yellowish-white cancellous bone pieces was all that remained in the original tube. Marrow cells were separated into a single cell suspension by passing them through syringes fitted with 18 and 20 gauge needles. Cells were spun at 1000×g for 5 minutes after which the fat layer and supernatant were removed. Cells were reconstituted in complete medium, counted with a hemocytometer (red blood cells were lysed prior to counting with 4% acetic acid), and plated in 100 mm dishes at 50×106 nucleated cells/dish.
Marrow for cell culture was prepared from aspirated bone marrow as follows. Aspirate marrow (5-10 ml) was transferred to sterile 50 ml plastic centrifuge tubes to which 20 ml complete medium was added. The tubes were spun at 1,500 rpm for 5 minutes to pellet the cells. The supernatant and fat layer were removed and the cell pellets were resuspended to 5 ml with complete medium. The cell suspensions then were loaded onto 70% Percoll (Sigma, St. Louis, Mo.) gradients with a 10 ml pipette and spun at 2,000 rpm (460×g) in a GS34 rotor for 15 minutes. In order to harvest the cells, the tube is marked just below the high concentrated band of platelets, about 25% to 35% of the way down the tube. (Pooled density=1.03g/ml.) Using a 10 ml pipette, the medium is aspirated off from the top down to the marked line (approximately 12-14 ml). the collected fraction is transferred to a 50 ml conical plastic tube. 30 ml of complete medium then is added to the tube and centrifuged for 5 minutes at 1,500 rpm. The supernatant then is removed and discarded.
The cells then are resuspended in 7 ml of complete medium, and a uniform cell suspension is generated by piperting cells up and down with a 10 ml pipette. The cells are counted by taking 20 μl of uniform cell suspension, and adding 20 μl of 4% acetic acid to the cell suspension. The suspension is mixed and transferred to a hemocytometer, and the cells are counted.
Cell concentration then is adjusted with complete medium to 50×106 cells per 7 ml. 7 ml of cells (50×106 cells) then are plated per 100 mm culture plate.
The marrow mesenchymal stem cells were purified and expanded as follows. Marrow cells from either the femoral head cancellous bone or the iliac aspirate were cultured in complete medium (i.e., DMEM-LG medium with 10% fetal bovine serum) at 37° C. in humidified atmosphere containing 95% air and 5% CO2. On day 3 after plating, nonadherent cells were removed from the cultures by aspirating the original medium from the plates and replacing the original medium with 7 ml of fresh complete medium. Subsequent medium changes were performed every 3 to 4 days. This process of removing the non-adherent cells during culture media changes results in purification of the mesenchymal stem cells which selectively adhere to the culture plates. When primary culture dishes became nearly confluent, the medium was removed and each plate rinsed with 7 ml of sterile Tyrode's solution. The cells were detached with 4 ml of 0.25% trypsin with 0.1 mM EDTA (GIBCO) for 5 minutes at 37° C. The action of trypsin was stopped with 2 ml of calf serum. The cells then were collected with a 10 ml pipette and centrifuged for 5 minutes at 1,500 rpm. The supernatant then is removed, and the cells resuspended from each plate in 21 ml complete medium, and a uniform cell suspension is generated by gently pipetting the cells up and down in the pipette. The cells from each culture then are counted and plated in three new 100 ml plates in 7 ml complete medium. The three new plates of cells are termed first passage cultures. First passage cells then were allowed to expand until they became near confluent, and were then replated as described above to create second passage cultures. At each passage, aliquots of cells were cryopreserved in 90% fetal bovine serum with 10% DMSO (freezing medium).
Mesenchymal stem cells were obtained from 10 ml of autologous human bone marrow collected either at the time of autologous bone marrow harvest, or during a routine diagnostic bone marrow examination, from the iliac crest. These mesenchymal stem cells were culture expanded and infused into 14 patients who had hematologic malignancies, but had no evidence of active malignancy, and had not received chemotherapy or radiation therapy for at least 4 weeks before collection of mesenchymal stem cells. Five patients each received 1 million autologous mesenchymal stem cells, five patients each were given 10 million mesenchymal stem cells, and four patients each received 50 million mesenchymal stem cells. Toxicity for each patient was determined according to the National Cancer Institute toxicity grading scale. The toxicity grading scale was the same as that based upon the common toxicity criteria for a Phase II trial of high-dose sequential chemotherapy and peripheral blood stem cell autologous transplantation as initial therapy for patients with poor prognosis non-Hodgkin's lymphoma. (National Cancer institute document, draft 4/94: E2493.) Toxicity was graded on a scale from 0 to 4. No Grade 2 or greater toxic effects were observed, and in two patients that received 10 million mesenchymal stem cells, there appeared to be an increase in bone marrow cellularity.
A breast cancer patient undergoes a diagnostic posterior iliac crest bone marrow aspiration and biopsy using a local anesthetic. A small portion (2 to 3 ml) of the aliquot (10 to 20 ml) of marrow is submitted for routine histologic testing and determination of tumor content using immunoperoxidase testing of cells grown in semisolid agar as described in Ross, et al., Blood., Vol. 82, pgs. 2605-2610 (1993). The remainder of the cells are cultured as hereinabove described. After at least 3 weeks in culture, samples of the mesenchymal stem cells are submitted for cell counting, cell viability (Trypan Blue), and cell surface immunophenotyping (testing with monoclonal antibodies SH2, SH3, and SH4). SH2 is produced by a hybridoma cell line assigned ATCC Accession No. HB10743. SH3 is produced by a hybridoma cell line assigned ATCC Accession No. HB10744. SH4 is produced by a hybridoma cell line assigned ATCC Accession No. HB10745. Samples also are submitted for detection of occult breast cancer.
The patient also undergoes placement of a pheresis central venous catheter, and receives subcutaneous injections of G-CSF (filgrastin) 10 μg/kg/day as described in Peters, et al., Blood, Vol. 81, pgs. 1709-1719 (1993); Chao, et al., Blood, Vol. 81, pgs. 2031-2035 (1993); Sheridan, et al., The Lancet, Vol. 2, pgs. 891-895 (1989); and Winter, et al., Blood, Vol. 82, pg. 293a (1993). G-CSF injections begin at least 3 days before the first pheresis is initiated. G-CSF therapy is withheld if the white blood cell count rises above 40,000/μL and is resumed once the white blood cell count drops to less than 20,000/μL.
If the patient is receiving only G-CSF as the vehicle for "mobilization" of peripheral blood progenitor cells, the patient must not have received chemotherapy within 4 weeks of the planned pheresis. If the patient has received both conventional chemotherapy and G-CSF treatment for mobilization, the patient must not have received chemotherapy within 10 days of the planned pheresis, and the white blood cell count must be at least 800/μL and the platelet count at least 30,000/μL.
Daily pheresis procedures are performed using a Cobe Spectra instrument (Cobe, Lakewood, Col.), and each cellular collection is cryopreserved using a controlled-rate liquid nitrogen freezer, until at least 15×108 mononuclear cells/kg are collected--(Lazarus, et al., Bone Marrow Transplant, Vol. 7, pgs. 241-246 (1991)). Each peripheral blood progenitor cell will be processed and cryopreserved according to previously published techniques. (Lazarus, et al., J. Clin, Oncol., Vol. 10, pgs, 1682-1689) (1992); Lazarus et al., (1991)).
Eight days before the patient is infused with the autologous peripheral blood progenitor cells, the patient receives chemotherapy over a period of 96 hours (4 days), with the following chemotherapy agents:
1. Cyclophosphamide in a total dosage of 6 g/m2 (1.5 g/m 2/day for 4 days) is given via continuous intravenous infusion at 500 mg/m2 in 1,000 ml normal saline every 8 hours.
2. Thiotepa in a total dosage of 500 mg/m2 /day for 4 days) is given via continuous intravenous infusion at 125 mg/2 in 1,000 ml normal saline every 24 hours.
3. Carboplatin in a total dosage of 1,800 mg/m2 (200 mg/m2 /day for 4 days) is given via continuous intravenous infusion at 200 mg/m2 in 1,000 ml of 5% dextrose in water every 24 hours.
The patient also receives 500 mg of mesna in 50 ml normal saline IV over 15 minutes every 4 hours for 6 days (144 hours), beginning with the first dose of cyclophosphamide.
At least 72 hours after the completion of the chemotherapy, the mesenchymal stem cells are harvested from tissue culture flasks. Cells are collected using bovine trypsin (which is deactivated with soybean trypsin inhibitor), suspended at 10×106 cells/ml in TC199 containing 1% human serum albumin (infusion medium), washed twice in infusion medium, resuspended at approximately 106 cells/ml, and injected slowly intravenously over 15 minutes to provide a total dosage of from 10 to about 5×106 cells.
The day after the patient receives the mesenchymal stem cells, the frozen autologous peripheral blood progenitor cells are removed from the liquid nitrogen refrigerator, transported to the patient in liquid nitrogen, submersed in a 37° C. to 40° C. sterile water bath, and infused rapidly intravenously without additional filtering or washing steps. GM-CSF in an amount of 250 μg/m2 then is given as a daily subcutaneous injection, beginning 3 hours after completion of the autologous blood progenitor cell infusion. The GM-CSF is given daily until the peripheral blood neutrophil count exceeds 1,000/μL for three consecutive days.
It is to be understood, however, that the scope of the present invention is not to be limited to the specific embodiments described above. The invention may be practiced other than as particularly described and still be within the scope of the accompanying claims.
|Cited Patent||Filing date||Publication date||Applicant||Title|
|US5226914 *||16 Nov 1990||13 Jul 1993||Caplan Arnold I||Method for treating connective tissue disorders|
|US5461034 *||18 Jun 1993||24 Oct 1995||Yissum Research Development Company Of The Hebrew University Of Jerusalem||Osteogenic growth polypeptides identified from regenerating bone marrow|
|1||*||Hard. Demonstration of the tumorigenicity of transformed rat kidney cell lines by intravenous allotransplantation in the neonate. Int. Cancer: 30, 197 202 (1982).|
|2||Hard. Demonstration of the tumorigenicity of transformed rat kidney cell-lines by intravenous allotransplantation in the neonate. Int. Cancer: 30, 197-202 (1982).|
|Citing Patent||Filing date||Publication date||Applicant||Title|
|US5942225 *||23 Jul 1997||24 Aug 1999||Case Western Reserve University||Lineage-directed induction of human mesenchymal stem cell differentiation|
|US6010696 *||25 Mar 1998||4 Jan 2000||Osiris Therapeutics, Inc.||Enhancing hematopoietic progenitor cell engraftment using mesenchymal stem cells|
|US6328960||12 Mar 1999||11 Dec 2001||Osiris Therapeutics, Inc.||Mesenchymal stem cells for prevention and treatment of immune responses in transplantation|
|US6645727||25 May 2001||11 Nov 2003||Stemcell Technologies Inc.||Antibody compositions for preparing enriched mesenchymal progenitor preparations|
|US6723131||28 Feb 2001||20 Apr 2004||The Cleveland Clinic Foundation||Composite bone marrow graft material with method and kit|
|US6930222||22 Aug 2001||16 Aug 2005||The Scripps Research Institute||In vivo animal model of human leukemia|
|US6936281||21 May 2001||30 Aug 2005||University Of South Florida||Human mesenchymal progenitor cell|
|US7049072||3 Oct 2002||23 May 2006||University Of South Florida||Gene expression analysis of pluri-differentiated mesenchymal progenitor cells and methods for diagnosing a leukemic disease state|
|US7098027 *||23 Dec 2002||29 Aug 2006||Renomedix Institute, Inc.||Method for remyelinating a demyelinized lesion due to injury in the brain or spinal cord|
|US7182781||1 Nov 2000||27 Feb 2007||Regeneration Technologies, Inc.||Cervical tapered dowel|
|US7255879||12 Aug 2003||14 Aug 2007||Anthrogenesis Corporation||Post-partum mammalian placenta, its use and placental stem cells therefrom|
|US7303769||10 Feb 2005||4 Dec 2007||University Of South Florida||Method for purifying pluri-differentiated mesenchymal progenitor cells|
|US7311904||13 Feb 2002||25 Dec 2007||Anthrogenesis Corporation||Tissue matrices comprising placental stem cells, and methods of making the same|
|US7311905||13 Feb 2003||25 Dec 2007||Anthrogenesis Corporation||Embryonic-like stem cells derived from post-partum mammalian placenta, and uses and methods of treatment using said cells|
|US7442390||9 Jul 2004||28 Oct 2008||University Of South Florida||Method for enhancing engraftment of cells using mesenchymal progenitor cells|
|US7468276||22 Jun 2004||23 Dec 2008||Anthrogenesis Corporation||Placental stem cells|
|US7470538||24 Jun 2004||30 Dec 2008||Case Western Reserve University||Cell-based therapies for ischemia|
|US7491388||4 Nov 1999||17 Feb 2009||Osiris Therapeutics, Inc.||Uses of fibroblasts or supernatants from fibroblasts for the suppression of immune responses in transplantation|
|US7682803||13 Oct 2006||23 Mar 2010||Anthrogenesis Corporation||Immunomodulation using placental stem cells|
|US7691415||4 Dec 2007||6 Apr 2010||University Of South Florida||Method for preventing, or reducing the severity of, graft-versus-host disease using pluri-differentiated mesenchymal progenitor cells|
|US7700090||28 Dec 2006||20 Apr 2010||Anthrogenesis Corporation||Co-culture of placental stem cells and stem cells from a second source|
|US7863045||18 Jul 2008||4 Jan 2011||Tigenix N.V.||Isolation of skeletal precursor cells|
|US7914779||29 Oct 2007||29 Mar 2011||Anthrogenesis Corporation||Tissue matrices comprising placental stem cells, and methods of making the same|
|US7976836||22 Dec 2008||12 Jul 2011||Anthrogenesis Corporation||Treatment of stroke using placental stem cells|
|US7993918||3 Aug 2007||9 Aug 2011||Anthrogenesis Corporation||Tumor suppression using placental stem cells|
|US8057788||28 Dec 2006||15 Nov 2011||Anthrogenesis Corporation||Placental stem cell populations|
|US8057789||31 Oct 2007||15 Nov 2011||Anthrogenesis Corporation||Placental stem cells derived from post-partum mammalian placenta, and uses and methods of treatment using said cells|
|US8057826||19 May 2008||15 Nov 2011||University Of South Florida||Methods for reducing graft-versus-host disease using pluri-differentiated mesenchymal progenitor cells|
|US8178084||8 Sep 2003||15 May 2012||Quy Biosciences Limited||Pharmaceutical kits comprising mesenchymal stem cells|
|US8202703||30 Jul 2010||19 Jun 2012||Anthrogenesis Corporation||Placental stem cell populations|
|US8216566||14 Jan 2010||10 Jul 2012||Anthrogenesis Corporation||Treatment of multiple sclerosis using placental stem cells|
|US8263065||29 Sep 2008||11 Sep 2012||Anthrogenesis Corporation||Tumor suppression using human placental perfusate and human placenta-derived intermediate natural killer cells|
|US8293223||30 Sep 2011||23 Oct 2012||Anthrogenesis Corporation||Treatment of organ injuries and burns using placental stem cells|
|US8334135 *||31 Jul 2003||18 Dec 2012||Yves Saint Laurent Parfums||Stem cells from adipose tissue, and differentiated cells from said cells|
|US8354370||16 Jun 2008||15 Jan 2013||Garnet Biotherapeutics, Inc.||Administering a biological composition or compositions isolated from self-renewing colony forming somatic cell growth medium to treat diseases and disorders|
|US8367409||19 Nov 2009||5 Feb 2013||Anthrogenesis Corporation||Amnion derived adherent cells|
|US8435788||24 Jun 2010||7 May 2013||Anthrogenesis Corporation||Tissue matrices comprising placental stem cells|
|US8455250||28 Dec 2006||4 Jun 2013||Anthrogenesis Corporation||Co-culture of placental stem cells and stem cells from a second source|
|US8460650||12 Feb 2008||11 Jun 2013||Anthrogenesis Corporation||Treatment of inflammatory diseases using placental stem cells|
|US8486696||10 Feb 2005||16 Jul 2013||Garnet Biotherapeutics, Inc.||Cell populations which co-express CD49c and CD90|
|US8545833||16 May 2011||1 Oct 2013||Anthrogenesis Corporation||Treatment of radiation injury using placental stem cells|
|US8562972||23 Oct 2007||22 Oct 2013||Anthrogenesis Corporation||Methods and compositions for treatment of bone defects with placental cell populations|
|US8562973||6 Apr 2011||22 Oct 2013||Anthrogenesis Corporation||Treatment of sarcoidosis using placental stem cells|
|US8580563||27 Oct 2008||12 Nov 2013||Anthrogenesis Corporation||Placental stem cells|
|US8586360||1 Jul 2010||19 Nov 2013||Anthrogenesis Corporation||Method of producing erythrocytes without feeder cells|
|US8591883||16 May 2012||26 Nov 2013||Anthrogenesis Corporation||Placental stem cell populations|
|US8603462||31 Mar 2004||10 Dec 2013||University Of Utah Research Foundation||Stem-cell, precursor cell, or target cell-based treatment of multi-organ failure and renal dysfunction|
|US8614191||2 Feb 2012||24 Dec 2013||Medipost Co., Ltd.||Use of ICAM-1 for prevention or treatment of neurological diseases|
|US8617535||3 Nov 2006||31 Dec 2013||Anthrogenesis Corporation||Cytotherapeutics, cytotherapeutic units and methods for treatments using them|
|US8691217||24 May 2012||8 Apr 2014||Anthrogenesis Corporation||Placental stem cell populations|
|US8728805||24 Aug 2009||20 May 2014||Anthrogenesis Corporation||Methods and compositions for treatment of bone defects with placental cell populations|
|US8753883||13 May 2011||17 Jun 2014||Anthrogenesis Corporation||Treatment of psoriasis using placental stem cells|
|US8828376||20 Aug 2009||9 Sep 2014||Anthrogenesis Corporation||Treatment of stroke using isolated placental cells|
|US8895256||17 May 2012||25 Nov 2014||Anthrogenesis Corporation||Immunomodulation using placental stem cells|
|US8916146||10 May 2013||23 Dec 2014||Anthrogenesis Corporation||Treatment of inflammatory diseases using placental stem cells|
|US8926964||13 Jul 2011||6 Jan 2015||Anthrogenesis Corporation||Methods of generating natural killer cells|
|US8969315||29 Dec 2011||3 Mar 2015||Anthrogenesis Corporation||Enhancement of placental stem cell potency using modulatory RNA molecules|
|US9040035||31 May 2012||26 May 2015||Anthrogenesis Corporation||Treatment of pain using placental stem cells|
|US9045735||3 Apr 2008||2 Jun 2015||The Cleveland Clinic Foundation||Enrichment of tissue-derived adult stem cells based on retained extracellular matrix material|
|US9078898||14 Feb 2014||14 Jul 2015||Anthrogenesis Corporation||Placental stem cell populations|
|US9115344||25 May 2012||25 Aug 2015||Nc Medical Research Inc.||Mesenchymal stem cell and a method of use thereof|
|US9121007||25 Jan 2011||1 Sep 2015||Anthrogenesis Corporatin||Treatment of bone-related cancers using placental stem cells|
|US9139813||15 Apr 2013||22 Sep 2015||Anthrogenesis Corporation||Renovation and repopulation of decellularized tissues and cadaveric organs by stem cells|
|US9149569||16 May 2011||6 Oct 2015||Anthrogenesis Corporation||Treatment of diseases or disorders using placental stem cells|
|US9198938||26 Dec 2012||1 Dec 2015||Antrhogenesis Corporation||Amnion derived adherent cells|
|US9200253||6 Aug 2008||1 Dec 2015||Anthrogenesis Corporation||Method of producing erythrocytes|
|US9216200||13 Aug 2012||22 Dec 2015||Anthrogenesis Corporation||Tumor suppression using human placental perfusate and human placenta-derived intermediate natural killer cells|
|US9254302||6 Apr 2011||9 Feb 2016||Anthrogenesis Corporation||Angiogenesis using placental stem cells|
|US9255248||11 Oct 2013||9 Feb 2016||Anthrogenesis Corporation||Method of producing erythrocytes without feeder cells|
|US9339520||16 Sep 2013||17 May 2016||Anthrogenesis Corporation||Methods and compositions for treatment of bone defects with placental cell populations|
|US9439931||19 Aug 2013||13 Sep 2016||Medipost Co., Ltd||Administering umbilical cord blood-derived mesenchymal stem cells to treat nerve injury|
|US9464274||18 Nov 2014||11 Oct 2016||Anthrogenesis Corporation||Methods of generating natural killer cells|
|US9539288||22 Oct 2014||10 Jan 2017||Anthrogenesis Corporation||Immunomodulation using placental stem cells|
|US9763983||4 Feb 2014||19 Sep 2017||Anthrogenesis Corporation||Natural killer cells from placenta|
|US9782441 *||10 Jul 2012||10 Oct 2017||Albert Einstein College Of Medicine, Inc.||Stromal cell therapy in treatment of radiation injury|
|US9788950||22 Apr 2016||17 Oct 2017||Vivex Biomedical, Inc.||Cohesive bone composition|
|US20020058025 *||20 Apr 2001||16 May 2002||Prockop Darwin J.||Stromal cell use|
|US20020160510 *||13 Feb 2002||31 Oct 2002||Hariri Robert J.||Renovation and repopulation of decellularized tissues and cadaveric organs by stem cells|
|US20030003084 *||21 May 2001||2 Jan 2003||Beerelli Seshi||Human mesenchymal progenitor cell|
|US20030032179 *||13 Feb 2002||13 Feb 2003||Hariri Robert J.||Post-partum mammalian placenta, its use and placental stem cells therefrom|
|US20030059414 *||21 Sep 2001||27 Mar 2003||Ho Tony W.||Cell populations which co-express CD49c and CD90|
|US20030114367 *||30 Sep 1999||19 Jun 2003||Yehuda Shoenfeld||Composition for the prevention and/or treatment of artherosclerosis|
|US20030180269 *||13 Feb 2003||25 Sep 2003||Hariri Robert J.||Embryonic-like stem cells derived from post-partum mammalian placenta, and uses and methods of treatment using said cells|
|US20030203483 *||3 Oct 2002||30 Oct 2003||Beerelli Seshi||Human mesenchymal progenitor cell|
|US20040058397 *||9 Sep 2003||25 Mar 2004||Thomas Terry E.||Novel antibody compositions for preparing enriched mesenchymal progenitor preparations|
|US20040101959 *||21 Nov 2002||27 May 2004||Olga Marko||Treatment of tissue with undifferentiated mesenchymal cells|
|US20040121461 *||23 Dec 2002||24 Jun 2004||Renomedix Institute Inc.||Cell fractions containing cells capable of differentiating into neural cells|
|US20040171147 *||25 Nov 2003||2 Sep 2004||Hariri Robert J.||Cytotherapeutics, cytotherapeutic units and methods for treatments using them|
|US20040208861 *||12 May 2004||21 Oct 2004||Philadelphia Health And Education Corporation||Stromal cell use|
|US20040229351 *||31 Jul 2003||18 Nov 2004||Anne-Marie Rodriguez||Stem cells from adipose tissue, and differentiated cells from said cells|
|US20050059147 *||9 Jul 2004||17 Mar 2005||Beerelli Seshi||Human mesenchymal progenitor cell|
|US20050069527 *||24 Jun 2004||31 Mar 2005||Case Western Reserve University||Cell-based therapies for ischemia|
|US20050142119 *||10 Feb 2005||30 Jun 2005||Beerelli Seshi||Human mesenchymal progenitor cell|
|US20050143420 *||2 Dec 2004||30 Jun 2005||Moutouh-De Parseval Laure||Methods and compositions for the treatment and management of hemoglobinopathy and anemia|
|US20050197283 *||15 Apr 2005||8 Sep 2005||Vascular Biogenics Ltd.||Compositions containing beta 2-glycoprotein I for the prevention and/or treatment of vascular disease|
|US20050233452 *||10 Feb 2005||20 Oct 2005||Neuronyx, Inc.||Cell populations which co-express CD49c and CD90|
|US20050282276 *||26 Jul 2005||22 Dec 2005||Renomedix Institute, Inc.||Cell fractions containing cells capable of differentiating into neural cells|
|US20060130852 *||8 Sep 2003||22 Jun 2006||Smith Roger Kenneth W||Pharmaceutical kits comprising mesenchymal stem cells|
|US20060154235 *||23 Aug 2005||13 Jul 2006||Takahiro Ochiya||Human hepatocyte-like cells and uses thereof|
|US20070053888 *||6 Nov 2006||8 Mar 2007||Hariri Robert J||Use of umbilical cord blood to treat individuals having a disease, disorder or condition|
|US20070178071 *||31 Mar 2004||2 Aug 2007||Christof Westenfelder||Stem-cell, precursor cell, or target cell-based treatment of multiorgan failure and renal dysfunction|
|US20070224177 *||7 Feb 2007||27 Sep 2007||Ho Tony W||Cell populations which co-express CD49c and CD90|
|US20070231309 *||19 Mar 2007||4 Oct 2007||Neuronyx, Inc.||Cell populations which co-express CD49c and CD90|
|US20070253931 *||10 Jan 2007||1 Nov 2007||Osiris Therapeutics, Inc.||Use of mesenchymal stem cells for treating genetic diseases and disorders|
|US20070264232 *||2 May 2007||15 Nov 2007||Neuronyx, Inc.||Cell populations which co-express CD49c and CD90|
|US20070275362 *||28 Dec 2006||29 Nov 2007||James Edinger||Placental stem cell populations|
|US20080032401 *||28 Dec 2006||7 Feb 2008||James Edinger||Placental stem cell populations|
|US20080044848 *||8 Jun 2007||21 Feb 2008||Heidaran Mohammad A||Placental niche and use thereof to culture stem cells|
|US20080060805 *||31 Oct 2007||13 Mar 2008||Cdx Gas, Llc||Method and system for accessing subterranean deposits from the surface and tools therefor|
|US20080089872 *||4 Dec 2007||17 Apr 2008||Beerelli Seshi||Method for purifying pluri-differentiated mesenchymal progenitor cells|
|US20080102058 *||22 May 2007||1 May 2008||Philadelphia Health And Education Corporation||Stromal cell use|
|US20080131966 *||29 Oct 2007||5 Jun 2008||Hariri Robert J||Renovation and repopulation of decellularized tissues and cadaveric organs by stem cells|
|US20080152624 *||3 Aug 2007||26 Jun 2008||Casper Paludan||Tumor suppression using placental stem cells|
|US20080152629 *||31 Oct 2007||26 Jun 2008||James Edinger||Placental stem cell populations|
|US20080206343 *||12 Feb 2008||28 Aug 2008||Edinger James W||Hepatocytes and Chondrocytes from Adherent Placental StemCells; And CD34+ ,CD45- Placental Stem Cell-Enriched Cell Populations|
|US20080213228 *||23 Oct 2007||4 Sep 2008||Anthrogenesis Corporation||Methods and Compositions for Treatment of Bone Defects with Placental Cell Populations|
|US20080241112 *||10 May 2005||2 Oct 2008||Christof Westenfelder||Therapy of Kidney Diseases and Multiorgan Failure with Mesenchymal Stem Cells and Mesenchymal Stem Cell Conditioned Media|
|US20080241246 *||15 Nov 2007||2 Oct 2008||Arteriocyte Inc.||Cell-based therapies for treating liver disease|
|US20080241870 *||22 Jul 2005||2 Oct 2008||National Centre For Cell Scineces||Composition For Creating an Artificial Bone Marrow Like Environment and Use Thereof|
|US20080248004 *||13 Mar 2008||9 Oct 2008||Renomedix Institute, Inc.||Cell fractions containing cells capable of differentiating into neural cells|
|US20080254007 *||19 May 2008||16 Oct 2008||Beerelli Seshi||Human Mesenchymal Progenitor Cell|
|US20080286249 *||5 Mar 2008||20 Nov 2008||Varney Timothy R||Use of mesenchymal stem cells for treating genetic diseases and disorders|
|US20080289395 *||21 Aug 2007||27 Nov 2008||Universal Scientific Industrial Co., Ltd.||Testing machine|
|US20090053183 *||16 Jun 2008||26 Feb 2009||Neuronyx Inc.||Treatment of Diseases and Disorders Using Self-Renewing Colony Forming Cells Cultured and Expanded In Vitro|
|US20090068207 *||11 Apr 2006||12 Mar 2009||Vascular Biogenics Ltd.||Compositions Containing Beta 2-Glycoprotein I-Derived Peptides for the Prevention and/or Treatment of Vascular Disease|
|US20090104164 *||26 Sep 2008||23 Apr 2009||Celgene Cellular Therapeutics||Angiogenic cells from human placental perfusate|
|US20090123927 *||18 Jul 2008||14 May 2009||Tigenix N.V.||Isolation of precursor cells and their use for tissue repair|
|US20090136471 *||7 Nov 2008||28 May 2009||Anthrogenesis Corporation||Treatment of premature birth complications|
|US20090142831 *||22 Dec 2008||4 Jun 2009||Anthrogenesis Corporation||Placental stem cells|
|US20090214487 *||11 Jan 2007||27 Aug 2009||Timothy Varney||Use of mesenchymal stem cells for treating genetic diseases and disorders|
|US20090215434 *||29 Apr 2009||27 Aug 2009||Cvon Innovations Limited||Method and system for distributing data to mobile devices|
|US20090252710 *||29 Sep 2008||8 Oct 2009||Celgene Cellular Therapeutics||Tumor suppression using human placental perfusate and human placenta-derived intermediate natural killer cells|
|US20090291061 *||21 May 2009||26 Nov 2009||Riordan Neil H||Stem cell therapy for blood vessel degeneration|
|US20100040588 *||10 Nov 2009||18 Feb 2010||Cryo-Cell International, Inc.||Procurement, Isolation, and Cryopreservation of Endometrial/Menstrual Cells|
|US20100047213 *||20 Aug 2009||25 Feb 2010||Andy Zeitlin||Cell composition and methods of making the same|
|US20100047214 *||24 Aug 2009||25 Feb 2010||Abramson Sascha D||Methods and Compositions for Treatment of Bone Defects with Placental Cell Populations|
|US20100047351 *||20 Aug 2009||25 Feb 2010||Andy Zeitlin||Treatment of stroke using isolated placental cells|
|US20100098671 *||7 Jun 2007||22 Apr 2010||Modern Cell & Tissue Technologies Inc.||composition for prevention or treatment of bone marrow damage|
|US20100098672 *||19 Oct 2009||22 Apr 2010||Timothy Varney||Use of mesenchymal stem cells for treating genetic diseases and disorders|
|US20100124569 *||19 Nov 2009||20 May 2010||Abbot Stewart||Amnion derived adherent cells|
|US20100143312 *||23 Nov 2009||10 Jun 2010||Hariri Robert J||Treatment of diseases, disorders or conditions of the lung using placental cells|
|US20100167316 *||3 Apr 2008||1 Jul 2010||The Cleveland Clinic Foundation||Enrichment of Tissue-Derived Adult Stem Cells Based on Retained Extracellular Matrix Material|
|US20100172830 *||8 Feb 2008||8 Jul 2010||Cellx Inc.||Extraembryonic Tissue cells and method of use thereof|
|US20100260847 *||24 Jun 2010||14 Oct 2010||Anthrogenesis Corporation||Tissue matrices comprising placental stem cells|
|US20100291047 *||28 Jul 2010||18 Nov 2010||Varney Timothy R||Use of Mesenchymal Stem Cells for Treating Genetic Disease and Disorders|
|US20100330052 *||2 Sep 2010||30 Dec 2010||Varney Timothy R||Use of Mesenchymal Stem Cells for Treating Genetic Diseases and Disorders|
|US20110003387 *||1 Jul 2010||6 Jan 2011||Abbot Stewart||Method of producing erythrocytes without feeder cells|
|US20110052670 *||1 Jul 2010||3 Mar 2011||University Of Southern California||Identification of a pluripotent pre-mesenchymal, pre-hematopoietic progenitor cell|
|US20110158960 *||3 Jan 2011||30 Jun 2011||Tigenix N.V.||Isolation of precursor cells and their use for tissue repair|
|US20110158967 *||4 Mar 2011||30 Jun 2011||NC MEDICAL RESEARCH INC. (75% interest)||Mesenchymal stem cell and the method of use thereof|
|US20110171659 *||19 Oct 2010||14 Jul 2011||Abt Holding Company||Multipotent Adult Stem Cells, Sources Thereof, Methods of Obtaining and Maintaining Same, Methods of Differentiation Thereof, Methods of Use Thereof and Cells Derived Thereof|
|US20110177045 *||31 Mar 2011||21 Jul 2011||Varney Timothy R||Use of Mesenchymal Stem Cells for Treating Genetic Disease and Disorders|
|US20110177595 *||7 Mar 2011||21 Jul 2011||Abt Holding Company||Multipotent Adult Stem Cells, Sources Thereof, Methods of Obtaining and Maintaining Same, Methods of Differentiation Thereof, Methods of Use Thereof and Cells Derived Thereof|
|US20110206645 *||25 Jan 2011||25 Aug 2011||Anthrogenesis Corporation||Treatment of bone-related cancers using placental stem cells|
|US20110217271 *||16 May 2011||8 Sep 2011||Anthrogenesis Corporation||Treatment of diseases or disorders using placental stem cells|
|US20110217272 *||16 May 2011||8 Sep 2011||Anthrogenesis Corporation||Treatment of radiation injury using placental stem cells|
|US20110223141 *||16 May 2011||15 Sep 2011||Anthrogenesis Corporation||Treatment of brain or spinal cord injury using placental stem cells|
|US20110262393 *||16 Nov 2009||27 Oct 2011||Medipost Co., Ltd.||Composition comprising mesenchymal stem cells or culture solution of mesenchymal stem cells for the prevention or treatment of neural diseases|
|US20140030235 *||26 Jul 2013||30 Jan 2014||Osiris Therapeutics, Inc.||Use of Mesenchymal Stem Cells for Treating Genetic Disease and Disorders|
|US20150118206 *||10 Jul 2012||30 Apr 2015||Chandan Guha||Stromal cell therapy in treatment of radiation injury|
|EP2204442A1||13 Oct 2008||7 Jul 2010||Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V.||Method of enriching stem cells in culture|
|EP2607477A1||5 May 2008||26 Jun 2013||The Brigham and Women's Hospital, Inc.||Multipotent stem cells and uses thereof|
|WO2001094541A2 *||21 May 2001||13 Dec 2001||University Of South Florida||Human mesenchymal progenitor cell|
|WO2001094541A3 *||21 May 2001||20 Jun 2002||Univ South Florida||Human mesenchymal progenitor cell|
|WO2002016559A2 *||21 Aug 2001||28 Feb 2002||The Scripps Research Institute||In vivo animal model of human leukemia|
|WO2002016559A3 *||21 Aug 2001||7 Aug 2003||Scripps Research Inst||In vivo animal model of human leukemia|
|WO2003029432A2 *||3 Oct 2002||10 Apr 2003||University Of South Florida||Human mesenchymal progenitor cell|
|WO2003029432A3 *||3 Oct 2002||15 Dec 2005||Univ South Florida||Human mesenchymal progenitor cell|
|WO2006092650A2 *||22 Jul 2005||8 Sep 2006||National Centre For Cell Sciences||A composition for creating an artificial bone -marrow like environment and use thereof|
|WO2006092650A3 *||22 Jul 2005||22 Feb 2007||Nat Ct Cell Sciences||A composition for creating an artificial bone -marrow like environment and use thereof|
|WO2013123094A2||13 Feb 2013||22 Aug 2013||The Regents Of The University Of California||Systemic delivery and regulated expression of paracrine genes for cardiovascular diseases and other conditions|
|U.S. Classification||424/93.7, 435/372, 424/423, 424/422|
|International Classification||A61F2/30, C07K16/28, C12N5/20, C12N5/00, A61L31/00, A61K35/12, A61K35/28, A61L27/38, A61F2/28, C12N5/077, C12N5/0775|
|Cooperative Classification||C12N2501/39, A61F2/30756, A61L2430/10, C07K16/28, A61K35/28, A61F2/28, A61L31/005, A61L27/3834, A61L27/3843, C12N2533/18, A61K2035/124, C12N5/0663, A61L27/3895, C12N2501/2301|
|European Classification||A61L27/38B14, A61L27/38H, A61L27/38D2, C12N5/06B13P1, C12N5/00S, A61K35/28, A61F2/28, C07K16/28, C12N5/06B21P, A61L31/00F|
|24 Jan 1995||AS||Assignment|
Owner name: CASE WESTERN RESERVE UNIVERSITY, OHIO
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:HAYNESWORTH, STEPHEN E.;CAPLAN, ARNOLD I.;GERSON, STANTON L.;AND OTHERS;REEL/FRAME:007326/0476
Effective date: 19950113
|9 Jun 1998||CC||Certificate of correction|
|15 Jul 1999||AS||Assignment|
Owner name: OSIRIS THERAPEUTICS, INC., MARYLAND
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:CASE WESTERN RESERVE UNIVERSITY;REEL/FRAME:010086/0736
Effective date: 19990706
|28 Sep 2001||FPAY||Fee payment|
Year of fee payment: 4
|30 Sep 2005||FPAY||Fee payment|
Year of fee payment: 8
|25 Sep 2009||FPAY||Fee payment|
Year of fee payment: 12
|1 Nov 2013||AS||Assignment|
Owner name: MESOBLAST INTERNATIONAL S?RL, SWITZERLAND
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:OSIRIS THERAPEUTICS, INC.;REEL/FRAME:031533/0828
Effective date: 20131010
Owner name: MESOBLAST INTERNATIONAL SARL, SWITZERLAND
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:OSIRIS THERAPEUTICS, INC.;REEL/FRAME:031533/0828
Effective date: 20131010